International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients

Schijman, Alejandro G.; Bisio, Margarita; Orellana, Liliana; Sued, Mariela; Duffy, Tomás; Mejia Jaramillo, Ana M.; Cura, Carolina; Auter, Frederic; Veron, Vincent; Qvarnstrom, Yvonne; Deborggraeve, Stijn; Hijar, Gisely; Zulantay, Inés; Lucero, Raúl Horacio; Velazquez, Elsa; Tellez, Tatiana; Sanchez Leon, Zunilda; Galvão, Lucia; Nolder, Debbie; Monje Rumi, María; Levi, José E.; Ramirez, Juan D.; Zorrilla, Pilar; Flores, María; Jercic, Maria I.; Crisante, Gladys; Añez, Néstor; Castro, Ana M. de; Gonzalez, Clara I.; Acosta Viana, Karla; Yachelini, Pedro; Torrico, Faustino; Robello, Carlos; Diosque, Patricio; Triana Chavez, Omar; Aznar, Christine; Russomando, Graciela; Büscher, Philippe; Assal, Azzedine; Guhl, Felipe; Sosa Estani, Sergio; Silva, Alexandre da; Britto, Constança; Luquetti, Alejandro; Ladzins, Janis

International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients

Detalhes bibliográficos
Autor(a) principal:	Schijman, Alejandro G.
Data de Publicação:	2011
Outros Autores:	Bisio, Margarita, Orellana, Liliana, Sued, Mariela, Duffy, Tomás, Mejia Jaramillo, Ana M., Cura, Carolina, Auter, Frederic, Veron, Vincent, Qvarnstrom, Yvonne, Deborggraeve, Stijn, Hijar, Gisely, Zulantay, Inés, Lucero, Raúl Horacio, Velazquez, Elsa, Tellez, Tatiana, Sanchez Leon, Zunilda, Galvão, Lucia, Nolder, Debbie, Monje Rumi, María, Levi, José E., Ramirez, Juan D., Zorrilla, Pilar, Flores, María, Jercic, Maria I., Crisante, Gladys, Añez, Néstor, Castro, Ana M. de, Gonzalez, Clara I., Acosta Viana, Karla, Yachelini, Pedro, Torrico, Faustino, Robello, Carlos, Diosque, Patricio, Triana Chavez, Omar, Aznar, Christine, Russomando, Graciela, Büscher, Philippe, Assal, Azzedine, Guhl, Felipe, Sosa Estani, Sergio, Silva, Alexandre da, Britto, Constança, Luquetti, Alejandro, Ladzins, Janis
Tipo de documento:	Artigo
Idioma:	eng
Título da fonte:	Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo:	https://www.arca.fiocruz.br/handle/icict/16262
Resumo:	Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET). Laboratorio de Biología Molecular de la Enfermedad de Chagas (LabMECh). Buenos Aires, Argentina.

Metadados do item

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oai_identifier_str	oai:www.arca.fiocruz.br:icict/16262
network_acronym_str	CRUZ
network_name_str	Repositório Institucional da FIOCRUZ (ARCA)
repository_id_str	2135
spelling	Schijman, Alejandro G.Bisio, MargaritaOrellana, LilianaSued, MarielaDuffy, TomásMejia Jaramillo, Ana M.Cura, CarolinaAuter, FredericVeron, VincentQvarnstrom, YvonneDeborggraeve, StijnHijar, GiselyZulantay, InésLucero, Raúl HoracioVelazquez, ElsaTellez, TatianaSanchez Leon, ZunildaGalvão, LuciaNolder, DebbieMonje Rumi, MaríaLevi, José E.Ramirez, Juan D.Zorrilla, PilarFlores, MaríaJercic, Maria I.Crisante, GladysAñez, NéstorCastro, Ana M. deGonzalez, Clara I.Acosta Viana, KarlaYachelini, PedroTorrico, FaustinoRobello, CarlosDiosque, PatricioTriana Chavez, OmarAznar, ChristineRussomando, GracielaBüscher, PhilippeAssal, AzzedineGuhl, FelipeSosa Estani, SergioSilva, Alexandre daBritto, ConstançaLuquetti, AlejandroLadzins, Janis2016-10-18T11:02:49Z2016-10-18T11:02:49Z2011SCHIJMAN, Alejandro G. et al. International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients. PLoS Negl Trop Dis, v. 5, n.1, e931, 13p,Jan. 2011.1935-2727https://www.arca.fiocruz.br/handle/icict/1626210.1371/journal.pntd.00009311935-2735engPublic Library of ScienceTrypanosoma cruziDoença de ChagasReação em Cadeia da PolimeraseMétodosAmostras de sangue humanoTrypanosoma cruziChagas DiseasePCR Methodshuman blood samplesInternational study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patientsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleInstituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET). Laboratorio de Biología Molecular de la Enfermedad de Chagas (LabMECh). Buenos Aires, Argentina.Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET). Laboratorio de Biología Molecular de la Enfermedad de Chagas (LabMECh). Buenos Aires, Argentina.Universidad de Buenos Aires (UBA). Instituto de Cálculo. Buenos Aires, Argentina.Universidad de Buenos Aires (UBA). Instituto de Cálculo. Buenos Aires, Argentina.Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET). Laboratorio de Biología Molecular de la Enfermedad de Chagas (LabMECh). Buenos Aires, Argentina.Universidad de Antioquia. Grupo Chagas. Medellin, Colombia.Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET). Laboratorio de Biología Molecular de la Enfermedad de Chagas (LabMECh). Buenos Aires, Argentina.French Blood Services. La Plaine Saint Denis, Paris, France.Universidad de Parasitología. Laboratorio Hospitalario. Cayene, French Guiana.Centers for Disease Control. Department of Parasitic Diseases. Atlanta, Georgia, USA.Institute of Tropical Medicine. Antwerp, Belgium.Instituto Nacional de Salud. Lima, Peru.Facultad de Medicina. Santiago de Chile, Chile.Universidad Nacional del Nordeste. Chaco, Argentina.Instituto Nacional de Chagas. Fatala Chabénn, Buenos Aires, Argentina.Universidad Mayor de San Simon. Facultad de Medicina. Centro Universitario de Medicina Tropical. Cochabamba, Bolivia.Universidad Nacional de Asunción. Instituto de Investigaciones en Ciencias de la Salud. Asunción del Paraguay, Paraguay.Faculdade de Farmácia. Natal, RN, Brasil.Hospital for Tropical Diseases. Department of Clinical Parasitology. London School of Tropical Medicine and Hygiene. London, United Kingdom.Universidad Nacional de Salta. Laboratorio de Patología Experimental. Salta, Argentina.Hospital Sirio Libanês. Banco de Sangue. São Paulo, SP, Brasil.Universidad de los Andes. Centro de Investigaciones en Microbiología y Parasitología Tropical. Bogotá, Colombia.Instituto Pasteur. Montevideo, Uruguay.Instituto de Salud Carlos III. Centro Nacional de Microbiologia. Centro de Mahahonda. Madrid, España.Instituto Nacional De Salud. Sección Parasitología. Santiago de Chile, Chile.Universidad de los Andes. Centro de Investigaciones Parasitológicas ‘‘J.F. Torrealba’’. Mérida, Venezuela.Universidad de los Andes. Centro de Investigaciones Parasitológicas ‘‘J.F. Torrealba’’. Mérida, Venezuela.Universidade Federal de Goiás. Instituto de Patologia Tropical e Saúde Pública (IPTSP). Goiânia, GO, Brasil.Universidad Industrial de Santander. Facultad de Salud. Grupo de Inmunologı´a y Epidemiologı´a Molecular (GIEM). Bucaramanaga, Colombia.Universidad Autónoma de Yucatán. Centro de Investigaciones Regionales (CIR) ‘‘Dr Hideyo Noguchi’. Laboratorio de Biología Celular. Departamento de Biomedicina de Enfermedades Infecciosas y Parasitarias. Yucatán, México.Universidad Católica de Santiago del Estero. Instituto de Biomedicina. Santiago del Estero, Argentina.Universidad Mayor de San Simon. Facultad de Medicina. Centro Universitario de Medicina Tropical. Cochabamba, Bolivia.Instituto Pasteur. Montevideo, Uruguay.Universidad Nacional de Salta. Laboratorio de Patología Experimental. Salta, Argentina.Universidad de Antioquia. Grupo Chagas. Medellin, Colombia.Universidad de Parasitología. Laboratorio Hospitalario. Cayene, French Guiana.Universidad Nacional de Asunción. Instituto de Investigaciones en Ciencias de la Salud. Asunción del Paraguay, Paraguay.Institute of Tropical Medicine. Antwerp, Belgium.French Blood Services. La Plaine Saint Denis, Paris, France.Universidad de los Andes. Centro de Investigaciones en Microbiología y Parasitología Tropical. Bogotá, Colombia.Centro Nacional de Diagnóstico e Investigación de Endemoepidemias (CeNDIE) ANLIS Dr. Carlos G. Malbrán. Buenos Aires, Argentina.Centers for Disease Control. Department of Parasitic Diseases. Atlanta, Georgia, USA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.Laboratório de Pesquisa de Doença de Chagas. Goiânia, GO, Brasil.World Health Organization (WHO). Special Programme for Research and Training in Tropical Diseases (TDR). Geneve, Switzerland.Background: A century after its discovery, Chagas disease still represents a major neglected tropical threat. Accurate diagnostics tools as well as surrogate markers of parasitological response to treatment are research priorities in the field. The purpose of this study was to evaluate the performance of PCR methods in detection of Trypanosoma cruzi DNA by an external quality evaluation. Methodology/Findings: An international collaborative study was launched by expert PCR laboratories from 16 countries. Currently used strategies were challenged against serial dilutions of purified DNA from stocks representing T. cruzi discrete typing units (DTU) I, IV and VI (set A), human blood spiked with parasite cells (set B) and Guanidine Hidrochloride-EDTA blood samples from 32 seropositive and 10 seronegative patients from Southern Cone countries (set C). Forty eight PCR tests were reported for set A and 44 for sets B and C; 28 targeted minicircle DNA (kDNA), 13 satellite DNA (Sat-DNA) and the remainder low copy number sequences. In set A, commercial master mixes and Sat-DNA Real Time PCR showed better specificity, but kDNA-PCR was more sensitive to detect DTU I DNA. In set B, commercial DNA extraction kits presented better specificity than solvent extraction protocols. Sat-DNA PCR tests had higher specificity, with sensitivities of 0.05–0.5 parasites/ mL whereas specific kDNA tests detected 5.1023 par/mL. Sixteen specific and coherent methods had a Good Performance in both sets A and B (10 fg/ml of DNA from all stocks, 5 par/mL spiked blood). The median values of sensitivities, specificities and accuracies obtained in testing the Set C samples with the 16 tests determined to be good performing by analyzing Sets A and B samples varied considerably. Out of them, four methods depicted the best performing parameters in all three sets of samples, detecting at least 10 fg/ml for each DNA stock, 0.5 par/mL and a sensitivity between 83.3–94.4%, specificity of 85–95%, accuracy of 86.8–89.5% and kappa index of 0.7–0.8 compared to consensus PCR reports of the 16 good performing tests and 63–69%, 100%, 71.4–76.2% and 0.4–0.5, respectively compared to serodiagnosis. Method LbD2 used solvent extraction followed by Sybr-Green based Real time PCR targeted to Sat-DNA; method LbD3 used solvent DNA extraction followed by conventional PCR targeted to Sat-DNA. The third method (LbF1) used glass fiber column based DNA extraction followed by TaqMan Real Time PCR targeted to Sat-DNA (cruzi 1/cruzi 2 and cruzi 3 TaqMan probe) and the fourth method (LbQ) used solvent DNA extraction followed by conventional hot-start PCR targeted to kDNA (primer pairs 121/122). These four methods were further evaluated at the coordinating laboratory in a subset of human blood samples, confirming the performance obtained by the participating laboratories. Conclusion/Significance: This study represents a first crucial step towards international validation of PCR procedures for detection of T. cruzi in human blood samples.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/16262/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALconstança_britto_etal_IOC_2011.pdfconstança_britto_etal_IOC_2011.pdfapplication/pdf885168https://www.arca.fiocruz.br/bitstream/icict/16262/2/constan%c3%a7a_britto_etal_IOC_2011.pdf899a7d486740a706f8e5aaeb05fb7dc4MD52TEXTconstança_britto_etal_IOC_2011.pdf.txtconstança_britto_etal_IOC_2011.pdf.txtExtracted texttext/plain72514https://www.arca.fiocruz.br/bitstream/icict/16262/3/constan%c3%a7a_britto_etal_IOC_2011.pdf.txt5b39489d11ee28d7cfa5eaaca45309f7MD53icict/162622018-04-02 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dc.title.pt_BR.fl_str_mv	International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients
title	International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients
spellingShingle	International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients Schijman, Alejandro G. Trypanosoma cruzi Doença de Chagas Reação em Cadeia da Polimerase Métodos Amostras de sangue humano Trypanosoma cruzi Chagas Disease PCR Methods human blood samples
title_short	International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients
title_full	International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients
title_fullStr	International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients
title_full_unstemmed	International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients
title_sort	International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients
author	Schijman, Alejandro G.
author_facet	Schijman, Alejandro G. Bisio, Margarita Orellana, Liliana Sued, Mariela Duffy, Tomás Mejia Jaramillo, Ana M. Cura, Carolina Auter, Frederic Veron, Vincent Qvarnstrom, Yvonne Deborggraeve, Stijn Hijar, Gisely Zulantay, Inés Lucero, Raúl Horacio Velazquez, Elsa Tellez, Tatiana Sanchez Leon, Zunilda Galvão, Lucia Nolder, Debbie Monje Rumi, María Levi, José E. Ramirez, Juan D. Zorrilla, Pilar Flores, María Jercic, Maria I. Crisante, Gladys Añez, Néstor Castro, Ana M. de Gonzalez, Clara I. Acosta Viana, Karla Yachelini, Pedro Torrico, Faustino Robello, Carlos Diosque, Patricio Triana Chavez, Omar Aznar, Christine Russomando, Graciela Büscher, Philippe Assal, Azzedine Guhl, Felipe Sosa Estani, Sergio Silva, Alexandre da Britto, Constança Luquetti, Alejandro Ladzins, Janis
author_role	author
author2	Bisio, Margarita Orellana, Liliana Sued, Mariela Duffy, Tomás Mejia Jaramillo, Ana M. Cura, Carolina Auter, Frederic Veron, Vincent Qvarnstrom, Yvonne Deborggraeve, Stijn Hijar, Gisely Zulantay, Inés Lucero, Raúl Horacio Velazquez, Elsa Tellez, Tatiana Sanchez Leon, Zunilda Galvão, Lucia Nolder, Debbie Monje Rumi, María Levi, José E. Ramirez, Juan D. Zorrilla, Pilar Flores, María Jercic, Maria I. Crisante, Gladys Añez, Néstor Castro, Ana M. de Gonzalez, Clara I. Acosta Viana, Karla Yachelini, Pedro Torrico, Faustino Robello, Carlos Diosque, Patricio Triana Chavez, Omar Aznar, Christine Russomando, Graciela Büscher, Philippe Assal, Azzedine Guhl, Felipe Sosa Estani, Sergio Silva, Alexandre da Britto, Constança Luquetti, Alejandro Ladzins, Janis
author2_role	author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author
dc.contributor.author.fl_str_mv	Schijman, Alejandro G. Bisio, Margarita Orellana, Liliana Sued, Mariela Duffy, Tomás Mejia Jaramillo, Ana M. Cura, Carolina Auter, Frederic Veron, Vincent Qvarnstrom, Yvonne Deborggraeve, Stijn Hijar, Gisely Zulantay, Inés Lucero, Raúl Horacio Velazquez, Elsa Tellez, Tatiana Sanchez Leon, Zunilda Galvão, Lucia Nolder, Debbie Monje Rumi, María Levi, José E. Ramirez, Juan D. Zorrilla, Pilar Flores, María Jercic, Maria I. Crisante, Gladys Añez, Néstor Castro, Ana M. de Gonzalez, Clara I. Acosta Viana, Karla Yachelini, Pedro Torrico, Faustino Robello, Carlos Diosque, Patricio Triana Chavez, Omar Aznar, Christine Russomando, Graciela Büscher, Philippe Assal, Azzedine Guhl, Felipe Sosa Estani, Sergio Silva, Alexandre da Britto, Constança Luquetti, Alejandro Ladzins, Janis
dc.subject.other.pt_BR.fl_str_mv	Trypanosoma cruzi Doença de Chagas Reação em Cadeia da Polimerase Métodos Amostras de sangue humano
topic	Trypanosoma cruzi Doença de Chagas Reação em Cadeia da Polimerase Métodos Amostras de sangue humano Trypanosoma cruzi Chagas Disease PCR Methods human blood samples
dc.subject.en.pt_BR.fl_str_mv	Trypanosoma cruzi Chagas Disease PCR Methods human blood samples
description	Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET). Laboratorio de Biología Molecular de la Enfermedad de Chagas (LabMECh). Buenos Aires, Argentina.
publishDate	2011
dc.date.issued.fl_str_mv	2011
dc.date.accessioned.fl_str_mv	2016-10-18T11:02:49Z
dc.date.available.fl_str_mv	2016-10-18T11:02:49Z
dc.type.status.fl_str_mv	info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv	info:eu-repo/semantics/article
format	article
status_str	publishedVersion
dc.identifier.citation.fl_str_mv	SCHIJMAN, Alejandro G. et al. International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients. PLoS Negl Trop Dis, v. 5, n.1, e931, 13p,Jan. 2011.
dc.identifier.uri.fl_str_mv	https://www.arca.fiocruz.br/handle/icict/16262
dc.identifier.issn.pt_BR.fl_str_mv	1935-2727
dc.identifier.doi.none.fl_str_mv	10.1371/journal.pntd.0000931
dc.identifier.eissn.none.fl_str_mv	1935-2735
identifier_str_mv	SCHIJMAN, Alejandro G. et al. International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients. PLoS Negl Trop Dis, v. 5, n.1, e931, 13p,Jan. 2011. 1935-2727 10.1371/journal.pntd.0000931 1935-2735
url	https://www.arca.fiocruz.br/handle/icict/16262
dc.language.iso.fl_str_mv	eng
language	eng
dc.rights.driver.fl_str_mv	info:eu-repo/semantics/openAccess
eu_rights_str_mv	openAccess
dc.publisher.none.fl_str_mv	Public Library of Science
publisher.none.fl_str_mv	Public Library of Science
dc.source.none.fl_str_mv	reponame:Repositório Institucional da FIOCRUZ (ARCA) instname:Fundação Oswaldo Cruz (FIOCRUZ) instacron:FIOCRUZ
instname_str	Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str	FIOCRUZ
institution	FIOCRUZ
reponame_str	Repositório Institucional da FIOCRUZ (ARCA)
collection	Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv	https://www.arca.fiocruz.br/bitstream/icict/16262/1/license.txt https://www.arca.fiocruz.br/bitstream/icict/16262/2/constan%c3%a7a_britto_etal_IOC_2011.pdf https://www.arca.fiocruz.br/bitstream/icict/16262/3/constan%c3%a7a_britto_etal_IOC_2011.pdf.txt
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repository.name.fl_str_mv	Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
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International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients

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